ERH基因对人膀胱癌T24和5637细胞迁移、侵袭的影响

Effect of ERH gene on migration and invasion of human bladder cancer cells T24 and 5637

目的研究基本同源物增强子(ERH)对人膀胱癌T24和5637细胞迁移和侵袭作用的影响。方法将人膀胱癌T24和5637细胞的ERH敲除后,采用细胞划痕实验、Transwell细胞迁移实验、Transwell细胞侵袭实验验证其迁移和侵袭功能变化,Western blot检测细胞迁移与侵袭相关蛋白的表达,并通过裸鼠尾静脉转移实验验证敲除ERH基因后在体内对膀胱癌细胞转移能力的影响。结果 (1)细胞划痕实验结果显示,敲除ERH基因后的人膀胱癌5637和T24细胞迁移率均明显降低(P <0. 05);(2)Transwell细胞迁移实验、Transwell细胞侵袭实验结果显示,敲除ERH基因后的人膀胱癌5637和T24细胞迁移细胞计数和侵袭细胞计数均明显减少(P <0. 05)。(3)Western blot实验结果显示,敲除ERH基因后的人膀胱癌5637细胞和T24细胞中E-Cadherin表达明显升高(P <0. 05),而Fibronectin、Twist、Vimentin、Snail2蛋白的表达明显降低(P <0. 05)。(4)裸鼠尾静脉转移实验显示,与ERH正常组相比,ERH敲除组小鼠肺转移灶明显减少。结论体外和体内实验均提示,ERH基因敲除影响人膀胱癌T24和5637细胞的迁移与侵袭。

Objective To study the effect of enhancer of rudimentary homolog ( ERH) gene on migration and invasion in human bladder cancer T24 and 5637 cells. Methods After knocking out the ERH gene of human bladder cancer T24 and 5637 cells, Wound healing assay, Trans well cell migration assay and Transwell cell invasion assay were used to verify the migration and invasion function. Cell migration related protein was detected by Western blot. Nude mouse tail vein transfer assay was used to study the metastasis ability of bladder cancer cells in vivo. Results (1) The Wound healing assay showed that there were significant differences in the migration cell counts of human bladder cancer 5637 and T24 ( P < 0. 05).(2) There were significant differences in migration and invasion cell counts of Transwell assay ( P < 0. 05 ).(3) Western blot showed that the expression of E-Cadherin in human bladder cancer 5637 cells and T24 cells was significantly increased (P < 0. 05) after knocking out ERH gene, while the expression of Fibronectin, Twist,Vimentin and Snail2 protein were significantly decreased ( P < 0. 05 ).(4)Nude mouse tail vein transfer assay showed that lung metastases were significantly reduced in the ERH knockout group compared with the normal ERH group. Conclusions Both in vitro and in vivo experiments suggest that ERH knockout affects the migration and invasion of human bladder cancer T24 and 5637 cells.